Description
Lysine malonylation is a protein posttranslational modification with potentially broad biological impact. We present a protocol to generate stable gene knockdown K562 cell lines through lentiviral infection of a CRISPRi system followed by lysine malonylation measurement using mass spectrometry (MS). We detail gRNA vector cloning, lentiviral infection, cell line purification, protein digestion, malonyllysine enrichment, desalting, and MS acquisition and analysis. This protocol can be applied for CRISPRa cell line generation and lysine malonylation profiling in tissues, with slight changes.
[doi:10.25345/C5M32NM76]
[dataset license: CC0 1.0 Universal (CC0 1.0)]
Keywords: Malonylation ; Data-independent acquisition (DIA) ; Post-translational modifications ; Lysine acetyltransferases ; Stable gene knockdown K562 cell lines
Contact
Principal Investigators:
(in alphabetical order)
|
Birgit Schilling, Buck Institute, USA
|
| Submitting User: |
JoannaBons
|
| Number of Files: |
|
| Total Size: |
|
| Spectra: |
|
| Subscribers: |
|
| |
|
Owner |
Conditions:
|
|
|
Biological Replicates:
|
|
|
Technical Replicates:
|
|
|
| |
| Identification Results |
Proteins (Human, Remapped):
|
|
|
Proteins (Reported):
|
|
|
Peptides:
|
|
|
Variant Peptides:
|
|
|
PSMs:
|
|
|
| |
Differential Proteins:
|
|
|
Quantified Proteins:
|
|
|
| |
Click here to queue conversion of this dataset's submitted spectrum files
to open formats (e.g. mzML). This process may take some time.
When complete, the converted files will be available in the "ccms_peak"
subdirectory of the dataset's FTP space (accessible via the "FTP Download"
link to the right).
Number of distinct conditions across all analyses (original submission and reanalyses)
associated with this dataset.
Distinct condition labels are counted across all files submitted in the "Metadata" category
having a "Condition" column in this dataset.
"N/A" means no results of this type were submitted.
Number of distinct biological replicates across all analyses (original submission and reanalyses)
associated with this dataset.
Distinct replicate labels are counted across all files submitted in the "Metadata" category
having a "BioReplicate" or "Replicate" column in this dataset.
"N/A" means no results of this type were submitted.
Number of distinct technical replicates across all analyses (original submission and reanalyses)
associated with this dataset.
The technical replicate count is defined as the maximum number of times any one distinct
combination of condition and biological replicate was analyzed across all files submitted in the
"Metadata" category. In the case of fractionated experiments, only the first fraction is
considered.
"N/A" means no results of this type were submitted.
Originally identified proteins that were automatically
remapped by MassIVE to proteins in the
SwissProt
human reference database.
"N/A" means no results of this type were submitted.
Number of distinct protein accessions reported across all analyses (original submission and
reanalyses) associated with this dataset.
"N/A" means no results of this type were submitted.
Number of distinct unmodified peptide sequences reported across all analyses (original
submission and reanalyses) associated with this dataset.
"N/A" means no results of this type were submitted.
Number of distinct peptide sequences (including modified variants or peptidoforms) reported
across all analyses (original submission and reanalyses) associated with this dataset.
"N/A" means no results of this type were submitted.
Total number of peptide-spectrum matches (i.e. spectrum identifications) reported across all
analyses (original submission and reanalyses) associated with this dataset.
"N/A" means no results of this type were submitted.
Number of distinct proteins quantified across all analyses (original submission and reanalyses)
associated with this dataset.
Distinct protein accessions are counted across all files submitted in the "Statistical Analysis
of Quantified Analytes" category having a "Protein" column in this dataset.
"N/A" means no results of this type were submitted.
Number of distinct proteins found to be differentially abundant in at least one comparison
across all analyses (original submission and reanalyses) associated with this dataset.
A protein is differentially abundant if its change in abundance across conditions is found
to be statistically significant with an adjusted p-value <= 0.05 and lists no issues associated
with statistical tests for differential abundance.
Distinct protein accessions are counted across all files submitted in the "Statistical Analysis
of Quantified Analytes" category having a "Protein" column in this dataset.
"N/A" means no results of this type were submitted.
This dataset may not contain all raw spectra data as originally deposited in PRIDE.
It has been imported to MassIVE for reanalysis purposes, so its spectra data here may
consist solely of processed peak lists suitable for reanalysis with most software.